Glycophorins A, B, and C: a family of sialoglycoproteins. Isolation and preliminary characterization of trypsin derived peptides

Abstract

Two new sialoglycoproteins, glycophorin B and glycophorin C, were isolated from erythrocyte membranes by extraction with lithium diiodosalicylate, partition in aqueous phenol, gel filtration in detergent, and preparative polyacrylamide gel electrophoresis in sodium dodecyl sulfate. The two proteins were characterized by amino acid and carbohydrate analysis, separation of tryptic peptides, and isolation and purification of the amino terminal glycopeptide from each polypeptide chain. Glycophorin B is found in two forms in electrophoretograms of normal erythrocyte membranes, corresponding to monomer and dimer, as has been similarly described for glycophorin A. By using antibodies to a carboxy terminal determinant of glycophorin A, and direct staining of gels with antibodies and 125I-protein A from Staph. aureus, as well as by two-dimensional immunoelectrophoreis, only the two known forms of glycophorin A are detectable. The data confirm and extend the notion that the sialoglycoproteins in human red cells are dimeric molecules which are either preformed in the membrane or which can readily be generated in vitro. Only glycophorin A and glycophorin C are sensitive to trypsin while in situ in the intact red blood cells.